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1 PCR- (Code No. FIK-101) TOYOBO CO., LTD. Life Science Department OSAKA JAPAN A4849K

2 [1] 1 [2] 2 [3] 2 [4] 2 [5] 2 [6] 4 [7] 6 [8] 7 [9] 8 0

3 [1]

4 [2] 2x PCR Master Mix 1ml x Primer Mix 1ml 1-20 UNG(Uracil-DNA Glycosylase 200 l 1-20 [3] 100bp 1.5ml 0.2mlPCR 20ml 1.5ml 95 12,000rpm MultiNA MultiNA DNA-1000 SYBR Gold Code:DNA SYBR Molecular Probes Inc. [4] inva VT1, VT2, VT2vha, VT2vhb, VT2vp1 ipah ipah [5] PCR PCR 2

5 PCR PCR dttp dutp DNA UNG UNG dutp PCR dttp DNA UNG PCR UNG UNG UNG / ( /) ipah 10 4 inva VT 10 6 InvA VT ipah InvA VT ipah 3

6 [6] A, B A 1) 1.5ml 50μl 2) 1-2 3) 10μl 50 B 1) 20ml 2.5ml 2) 2-3 3) 2-3 4) 50 5) 2-3 6) 0.5ml 1.5ml 1) 1.5ml 95 5min 2) 12,000rpm X 3 3) 2μl PCR 4 PCR PCR 1) 2) 3) 4) 18μl 5.6 μl 2x PCR Master Mix 10.0 μl 10x Primer Mix 2.0 μl UNG 0.4 μl 2.0 μl 20.0 μl 4

7 PCR UNG : 20, 10 95, 2 : 94, 10 : 57, : 68, 30 : 94, 10 : 55, : 68, 30 : 94, 10 : 53, : 68, MultiNA PCR DNA 100bp DNA Ladder DNA MultiNA bp 5 mv bp 7 mv bp 2mV bp 2 mv 5

8 (1) 2 (2) [7] 6

9 [8] O O157 cfu/g) 0 7x10 2 7x10 3 7x10 4 7x10 5 7x10 6 PCR cfu/g) 0 7 x x x x x10 6 PCR cfu/g) 0 4 x x x x x10 6 PCR 50 O157 cfu/g) x10 2 7x10 3 7x10 4 7x10 2 7x10 3 7x x x x10 4 7

10 [9] 10 UV inva, VT, ipah 10 UV 8

11 9

12 () TEL FAX order_lifescience@toyobo.jp () TEL FAX order_lifescience@toyobo.jp TEL FAX :00 12:00, 13:00 17:00 () tech_osaka@toyobo.jp URL

17-05 THUNDERBIRD SYBR qpcr Mix (Code No. QPS-201, QPS-201T) TOYOBO CO., LTD. Life Science Department OSAKA JAPAN A4196K

17-05 THUNDERBIRD SYBR qpcr Mix (Code No. QPS-201, QPS-201T) TOYOBO CO., LTD. Life Science Department OSAKA JAPAN A4196K 17-05 THUNDERBIRD SYBR qpcr Mix (Code No. QPS-201, QPS-201T) TOYOBO CO., LTD. Life Science Department OSAKA JAPAN A4196K - 18 - [1] 1 [2] 2 [3] 3 [4] 5 [5] : RNA cdna 13 [6] 14 [7] 17 LightCycler TM Idaho

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(2) (2) (3) (3) (5) ABI PRISM 7700 (5) Roche LightCycler TM (7) BioFlux LineGene (9) 1 RT-PCR (10) (12) (13) F. Hoffmann-La Roche Ltd. Roche Molecular 05-8 SYBR Green Realtime PCR Master Mix (Code No. QPK-201, QPK-201T) TOYOBO CO., LTD. Life Science Department OSAKA JAPAN A3314K (2) (2) (3) (3) (5) ABI PRISM 7700 (5) Roche LightCycler TM (7) BioFlux

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